CN109580584A - The preparation method of saliva diagnostic sensor and the application of saliva diagnostic sensor - Google Patents
The preparation method of saliva diagnostic sensor and the application of saliva diagnostic sensor Download PDFInfo
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Abstract
The present invention discloses a kind of preparation method of saliva diagnostic sensor and the application of saliva diagnostic sensor: graphene oxide solution is coated in the metal film surface for the substrate for being covered with metal film, it is dry, and obtain the substrate that surface is covered with graphene oxide membrane；Gained substrate is placed in hydrazine steam and is restored, the substrate that surface is covered with graphene film is obtained；The graphene film surface for being covered with the substrate of graphene film to gained surface coats metal ion solution, and in-situ reducing keeps metal nano material fixed on the surface of graphene, dry to exhale saliva to diagnose sensor to get described.Saliva diagnostic sensor of the present invention shows enhancing Raman scattering effect and stability with excellent, simple to the detection process of stomach cancer marker in saliva diagnosis, and expense is low, as a result reliably；Graphene not only carries metal nanoparticle, and the marker as gastric cancer in adsorption layer selective absorption enrichment saliva, therefore the present invention saliva diagnostic sensor mentioned and its stomach cancer marker diagnosis of gastric cancer has a extensive future in detection saliva.
Description
Technical field
The present invention relates to saliva diagnosis and clinical detection field, it is related to the preparation method of a kind of saliva diagnostic sensor and logical
It crosses the salivary sensors that the preparation method is prepared and is detecting the application in disease marker contained by saliva, especially detecting
Application in saliva in stomach cancer marker can achieve the purpose of early gastric caacer detection.
Background technique
When being detected in early stage cancer and obtaining medical treatment, the survival of cancer patient is significantly improved.For stomach
Cancer, is diagnosed as 5 annual survival rates of terminal illness patient about 10%, and is diagnosed as the patient of early stage disease and deposits within 5 years with 90%
Motility rate.However, current technology condition, most of patients with gastric cancer often suffer from terminal illness when being diagnosed.More early discovery cancer,
It is advantageous for the treatment of cancer.In addition to this, accurate gastric cancer parting helps to select correct operation plan, and reduces stomach
The important means of mortality of carcinoma.Therefore, it early diagnoses and accurately typing is always two kinds of premises that gastric cancer is cured.However,
Traditional gastroscope method is limited to that early gastric caacer clinical symptoms are unobvious, and process is loaded down with trivial details, brings huge psychosoma pain to patient,
It is not suitable for extensive, long-time follow-up investigations.Therefore, pass through particular cancers correlating markings in the body fluid such as detection blood, saliva
Object early diagnoses gastric cancer disease and the accurate parting of gastric cancer is most worthy and convenient method.
But for many main cancers, each gene in body fluid, there are sensitivity and specificity for protide marker not
The problem of foot.For example, for protide marker cal9-9, ca72-4 most common in patients with gastric cancer body fluid and carcinomebryonic antigen
(CEA) stomach neoplasm that can only detect about 15~50% either phase is down to about 2~11% for early stage disease.Although logical
Crossing very high-frequency detection can be improved the accuracy rate of diagnosis, but the collection of body fluid is inconvenient, and pretreatment process is complicated, difficult
To do high-frequency detection.If the marker of the quick and convenient repeatable detection of energy, by very good solution this problem.
Compared to body fluid, saliva, which has, to be facilitated multi collect and is easier to sample pre-treatments, with advances in technology and respectively
The appearance of kind high precision analysis instrument, scientists start the content expansion to various components and each component in saliva
Further analysis and research, the saliva diagnosis detection means novel as one kind start to be used to diagnose the illness.Therefore, pass through
The method of the particular cancers marker in saliva is detected particularly suitable for monitoring progression of disease, surgical operation can be tracked and controlled
It treats, the curative effect of radiation and chemotherapy.
Currently, the method for saliva diagnosis mainly includes chromatographic technique and mass spectrum direct detecting method.Both methods has
Higher analysis detection sensitivity and accuracy, but sample treatment time and effort consuming, environmental disturbances are big, it is difficult to it excludes, reproducibility
It is the deficiencies of bad, very high to mass detector requirement, analysis result can be interfered.Further, since chromatographic equipment is expensive, heavy, inspection
The survey period is long, cost is big, and detection method is difficult to standardize, and limits its application in clinical diagnosis.In recent years, saliva diagnoses
Sensor technology experienced in the past few years largely to be innovated and improves, the especially continuous research of nano material,
The saliva diagnostic sensor constructed by different nano materials improves rapid pace, simplification detection method, intuitive performance detection knot
Fruit has obtained the extensive concern of researchers in world wide.
In general, saliva diagnosis process comprising there are two, first is that marker is enriched with from saliva, second is that detecting these
Marker simultaneously carries out interpretation of result.Therefore, we detect the side of stomach cancer marker in saliva using a kind of saliva diagnostic sensor
Method.We select graphene as the marker in adsorption layer absorption saliva first, are subsequently used for Surface enhanced Raman scattering skill
The high sensitivity of art detects the marker of absorption.
Graphene is the two dimensional crystal being made of carbon atom, and carbon atom arrangement is as the monoatomic layer of graphite, because its is excellent
Good mechanical performance, thermodynamic behaviour and conductive capability is studied and is applied in many fields.Graphene is considered as
Plane polycyclic aromatic hydrocarbons (PAH) atomic crystal, each carbon atom are sp2Hydridization, and contribute electronically forming on a remaining p track
Big pi bond can be acted on by π-π and adsorb some substances, especially to there is organic molecule, therefore be particularly suitable for just in absorption richness
Collect substance in saliva.In addition, huge specific surface area, it also can be as a kind of fixed many kinds of substance of excellent carrier.The most
Important, graphene can enhance an Important Platform of Raman scattering application with action face.
Raman spectrum is widely used in analytical chemistry and pharmaceutical chemistry, can be lossless rapidly to being that substance is determined
Property and quantitative detection.However, the extremely weak Raman excitation efficiency of analyte limits always its broader applications, therefore how to increase
Add Raman signal, be always the research hotspot of researcher, and is greatly enhanced using surface enhanced Raman scattering effect
Raman signal, which is considered pole, application value.Surface enhanced Raman scattering effect refers to some metal good conductors in special preparation
In surface or colloidal sol, in excitation area, since the enhancing of sample surfaces or the electromagnetic field of near surface leads to the drawing of binding molecule
The phenomenon that graceful scattered signal is greatly enhanced than normal Raman scattered signal.Metal nano material, especially gold nano grain are table
Face enhancing Raman scattering is filled with new vitality, and Surface enhanced Raman scattering signal can amplify 10 at present15Times, this can realize list
Molecular Detection.Just because of the detection method of this high sensitivity, Surface enhanced Raman scattering is widely used in detection at present
Field, especially progress trace detection.
Summary of the invention
The object of the present invention is to provide the application of a kind of preparation method of saliva diagnostic sensor and liquid diagnostic sensor, benefits
Used in golden film surface building graphene and gold nano grain structure, by graphene film in-situ reducing obtain metal nano
Particle, while utilizing the dispersibility of graphene control metal nanoparticle, it is ensured that the increased efficiency of Raman, and can guarantee
Enough repeatability.On prepared saliva diagnostic sensor, graphene is for the gastric cancer in selective absorption detection saliva
The surface enhanced Raman scattering effect of marker, gold nano grain carries out Sensitive Detection to the marker of absorption, reaches to gastric cancer
The purpose of early diagnosis.
The purpose of the present invention is what is be achieved through the following technical solutions:
The present invention provides a kind of preparation method of saliva diagnostic sensor, includes the following steps: step 1, is being covered with metal film
The golden film face of substrate coats graphene oxide solution, dry, obtains the substrate that surface is covered with graphene oxide membrane；Step 2, by institute
Substrate is placed in hydrazine steam ambient and restores, obtain the substrate that surface is covered with graphene film；Step 3 is covered with graphite to gained surface
The graphene film surface matel coated solion of the substrate of alkene film, it is dry to get the saliva diagnostic sensor.
Preferably, in said step 1, the metal film with a thickness of 200~1000 nanometers.
Preferably, in said step 1, the type of the metal film includes golden film, copper film or nickel film etc..
Preferably, in said step 1, the material of the substrate includes ito glass, silicon wafer, simple glass etc..
Preferably, in said step 1, it is electroless plating or sputtering that substrate, which is covered with the mode of metal film,.
Preferably, in said step 1, the concentration of the graphene oxide solution is 0.25~1 grams per liter；The oxidation
The dosage of graphene solution specifically: 1~10 microlitre of each substrate.
Preferably, in said step 1, the mode of the coating is drop coating or spin coating.
Preferably, in step 2, the reduction carries out within the enclosed space；Hydrazine hydrate in the hydrazine steam ambient
Dosage is specially that the amount of every 100~1000 milliliters of enclosure space addition hydrazine hydrate is 50~200 milliliters.
Preferably, in step 2, the temperature of the reduction is 20~40 degrees Celsius, the time is 3~24 hours；The reduction
It can be carried out in insulating box；It also needs that gained substrate is dried after reduction.
Preferably, in step 3, the concentration of the metal ion solution is 1~5 mM every liter；The metal ion is molten
The dosage of liquid is that each gained substrate coats 1~5 microlitre；The mode of the coating is drop coating；The metal ion solution includes
Gold chloride solion, nitric acid silver ion solution, chloroplatinic acid solion etc..
Preferably, it also needs to be rinsed with water gained substrate in step 3, before the drying, removes unreacted metal ion.
Preferably, in the preparation method, the condition of the drying are as follows: spontaneously dried under room temperature, the room temperature specifically wraps
Including temperature is 25~30 degrees Celsius.
The present invention also provides the saliva diagnosis that the preparation method of the saliva diagnostic sensor described in one kind is prepared to pass
Application of the sensor in the disease marker contained by detection saliva.
Preferably, the disease marker is stomach cancer marker.
Disease mark is detected using the saliva diagnostic sensor that the preparation method of above-mentioned saliva diagnostic sensor is prepared
The method of will object is as follows: upper saliva sample is dripped on saliva diagnostic sensor surface, then standing adsorption applies Raman spectrum
Instrument detects saliva diagnostic sensor surface, finally by the spy in the Raman map generated to disease marker in saliva
It levies peak and carries out principal component analysis, observe it and fall in region, to carry out the classification of the disease marker contained in judgement saliva.It is excellent
Selection of land, the condition of the standing adsorption are 25~37 degrees Celsius, 1~3 hour.
The utility model has the advantages that graphene film after dry, is passed through by graphene oxide drop coating on golden film surface in the present invention
Hydrazine steam is adsorbed, then aoxidizes and generates through electronation.Graphene film is also used as Jenner's grain of rice, platinum nanoparticle, silver nanoparticle simultaneously
Particle provides growth site and stabilizer, makes the uniform particle diameter, intensive and uniformly of Jenner's grain of rice, platinum nanoparticle, silver nano-grain
It is dispersed in graphene film upper surface.The efficiency that the saliva diagnostic sensor that this method obtains has Raman to enhance, and have enough
Repeatability.When different saliva diagnostic sensor detection same samples, relative standard deviation as a result is 6.35%.It is being applied to stomach
When cancer diagnoses, is combined by the stomach cancer marker of the determining different physical conditions of correspondence carry out Raman detection first, available 14
A peak (as shown in Figure 4 A) relevant to stomach cancer marker in saliva can by obtaining Fig. 4 B to the progress PCA analysis of this 14 peak
To find that the stomach cancer marker combination of corresponding different physical conditions there are different distributed areas, illustrate that this 14 peaks can be used for distinguishing
Stomach cancer marker combination.Then, it is dissipated using by the stomach cancer marker in selective enrichment saliva, and conducive to surface-enhanced Raman
It penetrates effect and obtains the Raman spectrum in saliva in marker.The saliva Raman spectrum such as Fig. 4 C institute obtained using saliva diagnostic sensor
Show there is apparent peak to occur.14 in spectrogram Raman peaks relevant to stomach cancer marker are being selected, its peak area is taken to carry out
PCA result is as shown in Figure 4 D, obtains three different regions.As shown, healthy person (c '), early carcinoma of stomach patient (b ') and evening
Phase patients with gastric cancer (a ') mutually disjoints collection, can obviously distinguish.This explanation applies saliva diagnostic sensor of the invention, can examine
Measure the existing different region between early carcinoma of stomach patient, late gastric cancer patient and Healthy People.Therefore, when carry out diagnosis sentence
When other, as long as same analyze row PCA when detecting 14 peaks in obtained Raman map, observe it and fall in region, thus into
Row judges the health states in saliva collection source.The result shows that verifying in 10 verification samples, accuracy rate is greater than
90%。
Compared with prior art, the present invention have it is following the utility model has the advantages that
1, the saliva diagnostic sensor of this method preparation is easy to operate, low-cost, and in detecting saliva when marker, mistake
Journey is simple, and speed is fast, and accuracy is good.Traditional method based on phase chromatography-mass spectroscopy (GC-MS) needs to carry out loaded down with trivial details solid phase
The pretreatment process (absorption of solid phase microextraction and desorption process, each sample at least need 2 hours) of the complexity such as micro-extraction, and
When carrying out separation detection by gas-chromatography, the time is long (pillar is activated and separated, each sample at least 2 hours), analytic process
Difficulty is big, various consumptive materials it is at high cost, need profession personnel and expensive instrument.And the saliva diagnosis referred in the present invention passes
Sensor preparation process is simple, reproducible, can be also particularly suitable for detecting on a large scale with large scale preparation, yield rate about 80%.
It is demonstrated experimentally that equally 20 samples of detection with GC-MS and when solid phase microextraction at least need one week time, and with of the invention
Method, it is only necessary to be no more than 3-12 hours.By the verifying of the practical saliva to 3 groups of 18 people, the differentiation that 3 groups can be completed
It opens, reaches early carcinoma of stomach patient, the difference (as shown in Figure 4) of late gastric cancer patient and Healthy People.
2, in this method, prepared saliva diagnostic sensor Raman reinforcing effect is obvious, as shown in figure 3, can be light
Detect 10-6Mole every liter of rhodamine 6G (R6G) therefore can be used for marker in trace detection saliva.It is different in saliva
Marker combination, can indicate different diseases, therefore, the saliva diagnostic sensor that the present invention refers to can be not only used for examining
Disconnected gastric cancer is also used as a kind of new platform, especially not of the same race for detecting the marker for corresponding to various disease in saliva
This kind of existing disease long-term in patient body of the cancer of class, the metabolic system of meeting long lasting effect patient, and make in saliva
Marker presence steady in a long-term is conducive to the saliva diagnostic sensor that the present invention refers to and diagnoses to this kind of disease.
Detailed description of the invention
Upon reading the detailed description of non-limiting embodiments with reference to the following drawings, other feature of the invention,
Objects and advantages will become more apparent upon:
Fig. 1 is the electron microscope of saliva diagnostic sensor；Wherein,
A, the scanning electron microscope (SEM) photograph obtained after graphene oxide is dry；
B, the scanning electron microscope (SEM) photograph of the structure of the gold nano grain and graphene on saliva diagnostic sensor surface；
The transmission electron microscope picture of C graphene oxide；
The transmission electron microscope picture of the structure of the gold nano grain and graphene on D saliva diagnostic sensor surface；
Fig. 2 be X-ray spectrogram, in uv-visible absorption spectrum figure graphene oxide, graphene, saliva diagnostic sensor table
Surface layer figure；Wherein,
In Fig. 2A, three curves in X-ray spectrogram are respectively graphene oxide (a), graphene (b) and saliva diagnostic sensor
Superficial layer (c)；
In Fig. 2 B, three curves in uv-visible absorption spectrum figure are respectively graphene oxide (a), graphene (b) and saliva
Diagnostic sensor superficial layer (c)；
Fig. 3 is that the saliva diagnostic sensor being prepared increases ability to the Raman of R6G；
Wherein, a curve is the saliva diagnostic sensor detection 10 using preparation-4The raman spectrum that the R6G of M is obtained；
B curve is the saliva diagnostic sensor detection 10 using preparation-6The raman spectrum that the R6G of M is obtained；
C curve is the raman spectrum that solid R6G is measured in golden film；
D curve is the blank raman spectrum of saliva diagnostic sensor；
Fig. 4 is the saliva diagnostic sensor detection marker acquired results schematic diagram with preparation；Wherein,
A, 14 Raman peaks of 14 kinds of marker standard items mixtures；
B, principal component analysis result is carried out by the peak area for scheming 14 peaks that A is indicated；
C, using the saliva diagnostic sensor of preparation for detecting late gastric cancer patient, the simulation of early carcinoma of stomach patient and Healthy People
Saliva (a, b, c) and true saliva (a ', b ', c ')；
D, respectively to 15 14 simulated in saliva sample and 18 resulting Surface enhanced Raman spectroscopy of true saliva sample
Peak area carries out PCA analysis, respectively obtains a, three regions being clearly separated of b, c, while also obtaining a ', b ', c ' three are bright
Show separated region.
Specific embodiment
The present invention is described in detail combined with specific embodiments below.Following embodiment will be helpful to the technology of this field
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that the ordinary skill of this field
For personnel, without departing from the inventive concept of the premise, various modifications and improvements can be made.These belong to the present invention
Protection scope.
It is used as hydrazine vapor source using hydrazine hydrate (85%, v/v) in the present invention, hydrazine steam conduct can produce by the volatilization of hydrazine
Reducing agent is used in the present invention, and when graphene oxide uniform adsorption hydrazine steam, reduction process will not cause the rupture of film, and
Reduction effect is good good, it is even more important that the hydrazine of uniform adsorption makes the metal nanoparticle generated uniform in size, and distribution is equal
It is even.
Embodiment 1
Present embodiments provide for a kind of preparation methods of saliva diagnostic sensor, comprising the following steps:
Step 1 sputters upper 300nm golden film as substrate on the glass cleaned up.The graphite oxide of 2 microlitre of 0.25 grams per liter
Substrate surface of the alkene solution drop coating after cleaning；After natural drying, the graphene oxide membrane of formation is divided as shown in Figure 1A and C
It dissipates smooth.
Step 2 is placed in the enclosure space of 50 cubic centimetres of sizes, while being added in the peripheral region of substrate
100ml hydrazine hydrate solution (85%, v/v) is used as hydrazine vapor source, is placed in 37 DEG C of insulating box after sealing 12 hours；
Step 3 after taking-up, stands half an hour, 1 microlitre of chlorauric acid solution (5 mM) is added dropwise, spontaneously dries.Finally, small with water
Heart cleaning electrode obtains the saliva diagnostic sensor based on graphene and gold nano grain, is stored in N2In environment.Obtained biography
For sensor surfaces as shown in figure 1 shown in B and D, gold nano grain is uniform in size, is evenly distributed.Yield rate about 85%.
Measure during the preparation process, graphene oxide, graphene, saliva diagnostic sensor superficial layer X-ray spectrogram,
Uv-visible absorption spectrum figure is shown in Fig. 2；More specifically, the surface of Fig. 2A graphene oxide, graphene, saliva diagnostic sensor
The X-ray spectrogram of layer, a is graphene oxide, b is graphene, c is saliva diagnostic sensor superficial layer；Fig. 2 B is graphite oxide
Alkene, graphene, saliva diagnostic sensor superficial layer uv-visible absorption spectrum figure, wherein a is graphene oxide, b is
Graphene, c are saliva diagnostic sensor superficial layer.As a result illustrate that graphene oxide is successfully reduced into graphene, and
There is gold nano grain to be fixed on the surface of graphene.
Finally saliva diagnostic sensor obtained is shown in Fig. 3 to the Raman increase ability of rhodamine 6G (R6G) to this method,
In, a curve is the saliva diagnostic sensor detection 10 using preparation-4The raman spectrum that the R6G of M is obtained；B curve is using preparation
Saliva diagnostic sensor detection 10-6The raman spectrum that the R6G of M is obtained；C curve is the Raman that solid R6G is measured in golden film
Spectrogram；D curve is the blank raman spectrum of saliva diagnostic sensor；As shown in Figure 3, saliva diagnosis sensing prepared by the present invention
Device Raman reinforcing effect is obvious, can easily detect 10-6 moles every liter of rhodamine 6G (R6G).This illustrates preparation of the present invention
Saliva diagnostor have the ability to detect the marker molecules in saliva completely.
Drip upper 10 μ l saliva sample on the saliva diagnostic sensor surface, 37 DEG C standing adsorption 3 hours.It is utilized after taking-up
HORIBA LabRAM HR Evolution spectrophotometer is detected, it is 785nm that condition, which is laser wavelength, and 10% swashs
Optical power, the time of integration are 1 second, and cycle-index is 1 time.14 Raman peaks (being specifically shown in Fig. 4 A) is carried out in obtained spectrogram
Analysis, then PCA analysis (analysis result is shown in Fig. 4 B) is carried out, which region is the result observed fall within, to carry out judgement saliva
The health states in liquid collection source.The saliva Raman spectrum obtained using saliva diagnostic sensor is as shown in Figure 4 C, and PCA result is such as
Shown in Fig. 4 D, healthy person (c '), early carcinoma of stomach patient (b ') and late gastric cancer patient (a ') mutually disjoint collection, can obviously distinguish
It opens.Accuracy rate is tall and big in 90%.When different saliva diagnostic sensor detection same samples, relative standard deviation as a result is
6.35%。
Embodiment 2
Present embodiments provide for a kind of preparation methods of saliva diagnostic sensor, comprising the following steps:
Step 1 sputters upper 200nm golden film as substrate on the silicon wafer cleaned up.The graphite oxide of 2 microlitre of 0.5 grams per liter
Substrate surface of the alkene solution drop coating after cleaning；
Step 2 after natural drying, is placed in the space of 200 cubic centimetres of sizes, while being added in the peripheral region in substrate
Enter 1000ml hydrazine hydrate solution (85%, v/v) as hydrazine vapor source, is placed in after sealing in 37 DEG C of insulating box 12 hours.
Step 3 after taking-up, stands half an hour, 1 microlitre of platinum acid chloride solution (10 mM) is added dropwise, spontaneously dries.Finally,
It is carefully cleaned with water, obtains the saliva diagnostic sensor based on graphene and Pt nanoparticle.Yield rate is about 75%.
Drip upper 10 μ l saliva sample on the saliva diagnostic sensor surface, 37 DEG C standing adsorption 1 hour.It is utilized after taking-up
HORIBA LabRAM HR Evolution spectrophotometer is detected, it is 785nm that condition, which is laser wavelength, and 10% swashs
Optical power, the time of integration are 1 second, and cycle-index is 1 time.14 Raman peaks are analyzed in obtained spectrogram, then are carried out
PCA analysis, which region is the result observed fall within, to judge the health states in saliva collection source.As a result
Show that the saliva diagnostic sensor equally can obviously distinguish healthy person, early carcinoma of stomach patient and late gastric cancer patient.Accurately
Rate is tall and big in 80%.When different saliva diagnostic sensor detection same samples, relative standard deviation as a result is 8.82%.
Comparative example 1
This comparative example is the comparative example of embodiment 1, comparison place be, metal film with a thickness of 50nm.
As a result: experiment shows that only the golden film of 50nm is easy to appear crackle, and stability is poor, leads to the yield rate of sensor
Lower than 40%.
Comparative example 2
This comparative example is the comparative example of embodiment 1, and comparison place is, the concentration of the graphene oxide solution is 0.1 gram/
It rises；The dosage of the graphene oxide solution specifically: 15 microlitres of each substrate；
As a result: experiment shows that graphene oxide solution is easy to spread out, makes the area of graphene oxide membrane when being added dropwise 15 microlitres
Excessive, area is about 3 times or more be added dropwise in 1-10 microlitres.Lead to absorption enrichment marker deterioration of efficiency, the metal of carrying is received
The density of rice grain is small, and Raman, which enhances efficiency, to be reduced, and obtained Raman spectral peaks are weak, are unfavorable for analyzing and determining.
Comparative example 3
This comparative example is the comparative example of embodiment 1, and comparison place is, the concentration of the metal ion solution is 10 mMs every
It rises；The dosage of the metal ion solution is that each gained substrate coats 0.5 microlitre；
As a result: it tests and shows when the dosage of metal ion solution is that each gained substrate coats 0.5 microlitre, the metal of generation
Nano particle is seldom, and Raman, which enhances efficiency, to be reduced, and obtained Raman spectral peaks are weak, are unfavorable for analyzing and determining.At the same time, 0.5
Microlitre solion be unable to complete and spread out on graphene film, not only be easy to cause nano particle quantity small, also have size
Uneven phenomenon, this is also extremely unfavorable for surface enhanced Raman scattering effect.
Specific embodiments of the present invention are described above.It is to be appreciated that the invention is not limited to above-mentioned
Particular implementation, those skilled in the art can make various deformations or amendments within the scope of the claims, this not shadow
Ring substantive content of the invention.
Claims (10)
1. a kind of preparation method of saliva diagnostic sensor, which comprises the steps of:
Step 1 coats graphene oxide solution in the golden film face for the substrate for being covered with metal film, dry, obtains surface and is covered with oxidation stone
The substrate of black alkene film；
Gained substrate is placed in hydrazine steam ambient and restores by step 2, obtains the substrate that surface is covered with graphene film；
Step 3, the graphene film surface for being covered with the substrate of graphene film to gained surface coat metal ion solution, dry, punching
Wash the saliva diagnostic sensor to get graphene film surface equipped with metal nano material.
2. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in said step 1,
The metal film with a thickness of 200~1000 nanometers.
3. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in said step 1,
The concentration of the graphene oxide solution is 0.25~1 grams per liter；
The dosage of the graphene oxide solution specifically: 1~10 microlitre of each substrate.
4. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in said step 1,
The type of the metal film is golden film, copper film or nickel film；
The material of the substrate is ito glass, silicon wafer or simple glass.
5. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in the step 2,
The reduction carries out within the enclosed space；
The amount of hydrazine hydrate is added in the enclosure space that the dosage of hydrazine hydrate is every 100~1000 milliliters in the hydrazine steam ambient
It is 50~200 milliliters.
6. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in the step 2,
The temperature of the reduction is 20~40 degrees Celsius, the time is 3~24 hours.
7. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in the step 3,
The metal ion solution is gold chloride solion, silver ion solution and chloroplatinic acid solion.
8. the preparation method of saliva diagnostic sensor according to claim 1, which is characterized in that in the step 3,
The concentration of the metal ion solution is 1~5 mM every liter；The dosage of the metal ion solution are as follows: each gained substrate
1~5 microlitre of coating.
9. a kind of saliva that the preparation method by the described in any item saliva diagnostic sensors of claim 1 to 8 is prepared
Application of the diagnostic sensor in the disease marker contained by detection saliva.
10. application of the saliva diagnostic sensor according to claim 9 in the disease marker contained by detection saliva, special
Sign is that the disease marker is stomach cancer marker.
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| IT201900007214A1 (en) * | 2019-05-24 | 2020-11-24 | I R C C S Centro Neurolesi Bonino Pulejo | METHOD OF DIAGNOSIS FOR CELIAC DISEASE |
| WO2022180614A1 (en) | 2021-02-26 | 2022-09-01 | International Iberian Nanotechnology Laboratory (Inl) | Graphene-based malaria sensor, methods and uses thereof |
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| CN107703118A (en) * | 2017-11-14 | 2018-02-16 | 苏州纳葛诺斯生物科技有限公司 | Saliva diagnostic sensor, preparation method and its application in hepatitis C is detected |
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2018
- 2018-12-27 CN CN201811613254.5A patent/CN109580584A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105606585A (en) * | 2015-12-21 | 2016-05-25 | 上海交通大学 | Expiration sensor, as well as preparation method and application thereof |
| CN107192704A (en) * | 2017-07-19 | 2017-09-22 | 苏州纳葛诺斯生物科技有限公司 | Saliva diagnostic sensor and preparation method and application |
| CN107703118A (en) * | 2017-11-14 | 2018-02-16 | 苏州纳葛诺斯生物科技有限公司 | Saliva diagnostic sensor, preparation method and its application in hepatitis C is detected |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| IT201900007214A1 (en) * | 2019-05-24 | 2020-11-24 | I R C C S Centro Neurolesi Bonino Pulejo | METHOD OF DIAGNOSIS FOR CELIAC DISEASE |
| WO2020240399A1 (en) * | 2019-05-24 | 2020-12-03 | Irccs Centro Neurolesi "Bonino- Pulejo" | Celiac disease diagnosis method |
| CN110297082A (en) * | 2019-06-12 | 2019-10-01 | 杭州启创生物技术有限公司 | The method of the markers such as saliva uric acid specific detection and diagnosis gout |
| WO2022180614A1 (en) | 2021-02-26 | 2022-09-01 | International Iberian Nanotechnology Laboratory (Inl) | Graphene-based malaria sensor, methods and uses thereof |
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